APAgene GOLD™ kits

APAgene™ GOLD-RT Genome Walking Kits can now be shipped at room temperature !
International customers can save on shipping fees!

Our kit was featured in Molecular Biotechnology (D. Cullen et al., 11 January 2011 pp. 1-13) where it rated the best amongst our competitors and in Food Chemistry (MA Fraiture et al, 15 March 2014 pp 60-69) as an approach in identifying unauthorised GMOs

Applications

APAgene™ GOLD Genome Walking Kits are designed to rapidly (1 day!) and reliably amplify unknown genomic DNA using our patented APA technology. Also available, and on promotion, is our APAgene WALKING service.

APAgene™ GOLD is the ideal genomic tool because it allows you to carry out an entire range of molecular genetic experiments:

1. Isolate unknown genomic sequences flanking transgenes (including T-DNAs, gene traps, transposons), sequence-tagged sites (STSs) or expressed sequence tags (ESTs).
2. Isolate localized sequences flanking known sequences from large clones and genomic DNAs.
3. Sequence insert ends of large clones such as P1, YAC and BAC DNAs.
4. Isolate unknown 5' promoter control regions and 3' transcriptional terminators of cDNAs.
5. Isolate unknown 5' and 3' RACE from first-strand cDNAs.

Amplification process

Our patented technology relies on a combination of our degenerate random tagging primers (DRT) and customer-provided gene-specific primers (GSP). Typically, three nested gene specific primers (GSPa, GSPb, and GSPc) are required in the reaction for the amplification of flanking sequences.

DRT primers are universal binding primers consisting of 3 parts:

1. a degenerate sequence
2. a random sequence
3. a tagging sequence

All the DRT primers share the same tagging sequence, but have different random sequences. The success of the walking reaction relies on the successful binding of the DRT primers to the flanking sequence during the primary PCR. Our patented DRT primers, along with our optimized cycling conditions and PCR buffer, greatly enhance the amplification process.

Offered kits

We offer 2 different APAgene™ GOLD Genome Walking Kits. Both kits yield excellent results. BT901-RT is our newest kit and the one we recommend to all. BT501 is an older version that many of our loyal clients prefer.Our new APAgene™ GOLD-RT Genome Walking Kits can be shipped at ROOM TEMPERATURE! The advantage is that shipping fees are reduced to a minimum compared to the significant dry ice shipping fees. For example, shipping to the UK would cost about $45 US.

BT901-RT (10 walk format)

APAgene™ GOLD-RT Genome Walking Kitsroom temperature for shipping. This kit utilizes UAP primers which are hairpin-containing primers making non-specific amplification much less efficient compared to targeted sequences.

A few references

• A naturally occurring epigenetic mutation in a gene encoding an SBP-box transcription factor inhibits tomato fruit ripening, Kenneth Manning et al., Nature Genetics, Volume: 38, Issue: 8, Date: 2006 07 28, Pages: 948-522
• A Na+: H+ Antiporter and a Molybdate Transporter Are Essential for Arsenite Oxidation in Agrobacterium tumefaciens, Journal of Bacteriology, February 2006, p. 1577-1584, Vol. 188, No. 4, Des R. Kashyap et al.
• Y chromosome microsatellite isolation from BAC clones in the greater white-toothed shrew (Crocidura russula), Lawson Handley, L.J., Perrin, N., 2006, Molecular Ecology Notes 60, 276-279.
• Complex Regulation of Arsenite Oxidation in Agrobacterium tumefaciens, Journal of Bacteriology, February 2006, p. 1081-1088, Vol. 188, No. 3, Des R. Kashyap et al.
• Identification and regulatory analysis of rainbow trout tapasin and tapasin-related genes, Immunogenetics 2006 Feb; 58(1) :56-69. Landis ED, Palti Y, Dekoning J, et al.
• Detection and Characterization of Amplified Fragment Length Polymorphism Markers for Clinical Mastitis in Canadian Holsteins, J Dairy Sci. 2006 Sep;89 (9):3653-63., B. S. Sharma et al.
• Homologs of CD83 from Elasmobranch and Teleost Fish, Yuko Ohta et al., The Journal of Immunology, 2004, 173: 4553-4560.